Characterization of carfilzomib-resistant non-small cell lung cancer cell lines View Full Text


Ontology type: schema:ScholarlyArticle      Open Access: True


Article Info

DATE

2018-05-15

AUTHORS

Neale T. Hanke, Elliot Imler, Marilyn T. Marron, Bruce E. Seligmann, Linda L. Garland, Amanda F. Baker

ABSTRACT

PurposeWe previously showed that carfilzomib (CFZ) has potent anti-proliferative and cytotoxic activity in a broad range of lung cancer cell lines. Here we investigate possible mechanisms of CFZ acquired resistance in lung cancer cell lines.MethodsCFZ-resistant non-small cell lung cancer (NSCLC) cell lines were developed by exposing A549 and H520 cells to stepwise increasing concentrations of CFZ. Resistance to CFZ and cross-resistance to bortezomib and other chemotherapy drugs was measured using the MTT assay. Cytotoxicity to CFZ was determined using a CytoTox assay. Western blot was used to measure apoptosis, autophagy, and drug efflux transporter-related proteins. Quantitative targeted whole transcriptome sequencing and quantitative RT-PCR was used to measure gene expression. Flow cytometry was used to analyze intracellular accumulation of doxorubicin.ResultsThe CFZ IC50 value of the resistant cells increased versus parental lines (2.5-fold for A549, 122-fold for H520). Resistant lines showed reduced expression of apoptosis and autophagy markers and reduced death versus parental lines following CFZ treatment. Both resistant lines exhibited higher P-glycoprotein (Pgp) gene (TempO-Seq® analysis, increased 1.2-fold in A549, > 9000-fold in H520) and protein expression levels versus parental lines. TempO-Seq® analysis indicated other drug resistance pathways were upregulated. The resistant cell lines demonstrated less accumulation of intracellular doxorubicin, and were cross-resistant to other Pgp client drugs: bortezomib, doxorubicin, and paclitaxel, but not cisplatin.ConclusionsUpregulation of Pgp appears to be an important, but not the only, mechanism of CFZ resistance in NSCLC cell lines. More... »

PAGES

1317-1327

Identifiers

URI

http://scigraph.springernature.com/pub.10.1007/s00432-018-2662-0

DOI

http://dx.doi.org/10.1007/s00432-018-2662-0

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1103994980

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/29766327


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